The movie begins at a blood bank. A homeless man walks into the clinic and sits by another homeless man (Luke Goss) with a scar running down his chin. He sees that the scarred man looks nervous and tries to relax him by telling him that they pay well there. The other man still looks uneasy as the nurse takes him into the clinic. They walk down a very dark and deserted hallway until they reach a room with several people waiting. The man starts to panic and is forced into the medical chair in the center of the room. One of the people starts a blood draining device and is about to suck out the homeless man's blood when the man's whimpering turned into laughter. He grabs one of the guards and drains her blood. He shows unbelievable strength as he kills everyone in the room. He looks at the surveillance camera and speaks in the vampire language, "I hate vampires."

After the prologue, we hear a narration explaining the events following the first movie. Even though Whistler shot himself in the first film. The vampires got to him before he died and revived him. Now Blade (Wesley Snipes) has been busy trying to find him so he can rest in peace. We see Blade chasing a bunch of vampires down an alley. In an elaborate battle sequence, he manages to kill all the vampires except for a pudgy one. He manages to find out where they were storing Whistler, and goes to free him. He lets the pudgy vampire go, saying "I'll catch you later." (IMPORTANT LATER) After another long battle, he does find Whistler (Kris Kristofferson). He was being suspended in a vat of blood. Blade releases him, and then takes him back to his headquarters. Blade has a new partner, Scud (Norman Reedus), who does not trust Whistler and tells Blade to kill him. Blade says that there is still a chance, since they kept him suspended all that time. He injects Whistler with an anti-virus and let him go cold turkey for a night.

The next day, Whistler has reverted back to human. They were just catching up when the alarms were tripped. Blade fights two masked vampires to a stalemate. One vampire stops the fight and said they were there to present a truce. They unmask themselves and one of the fighters is Nyssa (Leonor Varela), the daughter of the leader of the vampire nation.They take him to the fortress of the Vampire Leader, Damaskinos. The leader looks more like a demon than the other vampires, I guess because he is very old.He tells Blade that the vampire virus has mutated, and created a new race called Reapers. Reapers have a stronger craving for blood than vampires, and they will feed even off of vampires. When vampires are bitten, they will turn into Reapers. They isolated the spread of the virus to one carrier, which was the man on surveillance camera in the prologue. He is called Nomak.

Next, Blade is introduced to the Bloodpack. A group of elite vampires training for two years to hunt and kill Blade. Blade now leads them to hunt the Reapers. The group consists of Nyssa and the other vampire Blade fought earlier, plus Rienhardt (Ron Perlman), Chupa (Matt Schulze), Snowman (Donnie Yen), Lighthammer (Daz Crawford), Priest (Tony Curran), Verlaine (Marit Velle Kile). Rienhardt mouths off at Blade, so Blade attaches a small explosive on the back of his skull to keep him in line. Blade suggest they patrol vampire hangouts to find the Reapers.

The pack heads to a major vampire club, much like the one in the beginning of the first movie, but much bigger. They do find a group of Reapers. A full scale battle takes place. The Reapers are immune to bullets, silver, and garlic. Also they are extremely fast, strong, and durable and seem to feel no pain. When they are ready to bite, they can loosen their entire lower jaw (like the Predator). That is why they have a scar running down the lower jaw. Priest is killed, and Lighthammer is bitten. But he hides his wound. Nyssa is grabbed by Nomak, but he does not kill her (IMPORTANT LATER). Blade finds Nomak with Nyssa and shoots Nomak in the head, but there is no effect. In the mean time, Scud is attacked outside the club by Reapers. He turns on the ultraviolet light on his van, which kills the Reapers. It is the one weakness Reapers share with vampires. Scud tells the rest of the team about the weakness and they manage to wipe out some Reapers using ultraviolet lights on their guns. Blade fights Nomak hand to hand, but it is a draw. Nomak runs away due to the fact that sunlight was coming through the windows.

The pack manage to capture one Reaper. They examined him and realized that they need to feed all the time because of their high metabolism, or else they will die really fast. Only Nomak does not have this weakness because he is the carrier. Nyssa dissects the Reaper, and discovers that the heart was incased in bone. That was why they were so tough to kill. Scud decides to create an ultraviolet flashbang, since that is the only effective weapon they are aware of. Blade decides to take the pack out during day time, since the Reapers will be more vulnerable then. They cover themselves with pheromones extracted from the Reaper to attract them.

The pack head into the sewers, where they saw the Reapers head to after the club. They split up into teams. (Here the story diverge a little, so I will talk about each team a little out of order from the film sequence). When Blade is away, Chupa beats up Whistler, saying that he is going to pay for Priest's death. Whistler releases a batch pheromones to attract the Reapers. They overwhelm Chupa and feed off him while Whistler slips away. He runs into Nomak, who says he will let him live to deliver a message to Blade. He gives Whistler a ring and whisper something in his ear. Meanwhile, Lighthammer turns into a Reaper from his bite and kills Snowman. He chases after Verlaine, who tries to escape to the surface. Since it is day, when she opens the manhole cover, the light kills her and Reaper Lighthammer.

Finally, Blade is with Nyssa and the other one from the earlier fight. The other vampire and Nyssa are ambushed and the vampire is killed. Blade saves Nyssa and they run through the tunnels shooting at the horde of Reapers crawling out of every opening. They meet up with Rienhardt and form a circle just shooting at the Reapers that are slowly surrounding them. Blade tells Nyssa and Rienhardt to escape through a opening in the Reapers attack. He then release a ultraviolet flashbang, killing all the Reapers around him. He runs down a tunnel, to where the pack at previously placed a super flashbang. Blade sets it off, filling the entire tunnel with ultraviolet light, killing all the Reapers. Reinhardt and Nyssa escape most of the blast by diving under the water, but are still badly burned. Blade finds Nyssa, and feeds her some of his blood to save her. While he was doing that, he is hit by tasers and knocked out. Whistler is also captured.

Blade, Whistler, and Scud are back in the vampire leader's headquarters. We learn that the Leader was manipulating DNA to get rid of the weaknesses of the vampire virus, such as ultraviolet, garlic, and silver. Nomak was created in the experimentation. Whistler shows the ring to Nyssa, and she realized the crest on the ring was her family's. Nomak was her brother. Reinhardt shoots Blade in the leg. Blade uses his remote detonator, but it didn't explode. Turns out Scud was Reinhardt's Familiar (human lackey) and he gave Blade a dud. Reinhardt pulls off the explosive and throws it to Scud. Scud says he rather be a pet than prey. Blade laughs and says he knew Scud was working for Reinhardt. He says the bomb wasn't a dud and presses another button. Scud blows up. Henchmen shocks Blade with tasers and take him into a medical chamber. Blade is impaled onto a medical table. They are going to dissect him so they can use his physiology to perfect the experiments.

What they didn't count on was that Nomak had followed them back. He breaks into the headquarters and starts wrecking havoc. Whistler takes the opportunity to rescue Blade. Blade is weakened from losing so much blood, so Whistler dumps him in a pool of blood the leader uses to rejuvenate himself. Blade emerges with a strong bloodlust, like in the first movie. He beats up a bunch of henchmen, then splits Reinhardt into two. Meanwhile, the leader and Nyssa are escaping from Nomak. Suddenly, Nyssa lockdown the entire building. She reveals that Nomak had told her everything back at the club, and she was disgusted by her father's behavior. Nomak bites his father, but does not feed. He lets him bleed to death. He then feeds off Nyssa. Blade jumps in and they fight one on one.In the end, Blade manages to slip his sword between slits of the bone encasing Nomak's heart. Nomak sinks to the floor, and says it doesn't hurt anymore. He pushes the blade in by himself and disintegrates. Blade goes to Nyssa, who says she feels the change, and she wants to die as a vampire. She says she always wanted to see the sun. Blade takes her outside as dawn is coming on. Nyssa sees the sun and dies.

In the epilogue, we see the pudgy vampire from the beginning of the movie go into a sleazy stripper joint. He sits in a private booth facing a window. The light comes on the other side and it is Blade. He says "you didn't think I forgot about you did you?" He stabs his sword through the window and into the vampire's head.

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